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Optimization of vapor diffusion conditions for anti-CD20 crystallization and scale-up to meso batch

DOI: 10.3390/cryst9050230 DOI Help

Authors: Huaiyu Yang (Imperial College London) , Benny Danilo Belviso (Consiglio Nazionale delle Ricerche) , Xiaoyu Li (Imperial College London) , Wenqian Chen (Imperial College London) , Teresa Fina Mastropietro (Istituto per la Tecnologia delle Membrane, CNR) , Gianluca Di Profio (Istituto per la Tecnologia delle Membrane, CNR) , Rocco Caliandro (Istituto di Cristallografia, CNR) , Jerry Y. Y. Heng (Imperial College London)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Crystals , VOL 9

State: Published (Approved)
Published: April 2019
Diamond Proposal Number(s): 15832

Open Access Open Access

Abstract: The crystal form is one of the preferred formulations for biotherapeutics, especially thanks to its ability to ensure high stability of the active ingredient. In addition, crystallization allows the recovery of a very pure drug, thus facilitating the manufacturing process. However, in many cases, crystallization is not trivial, and other formulations, such as the concentrate solution, represent the only choice. This is the case of anti-cluster of differentiation 20 (anti-CD20), which is one of the most sold antibodies for therapeutic uses. Here, we propose a set of optimized crystallization conditions for producing anti-CD20 needle-shaped crystals within 24 h in a very reproducible manner with high yield. High crystallization yield was obtained with high reproducibility using both hanging drop vapor diffusion and meso batch, which is a major step forward toward further scaling up the crystallization of anti-CD20. The influence of anti-CD20 storage conditions and the effect of different ions on the crystallization processes were also assessed. The crystal quality and the high yield allowed the first crystallographic investigation on anti-CD20, which positively confirmed the presence of the antibody in the crystals.

Journal Keywords: anti-CD20; crystallization; X-ray diffraction

Subject Areas: Chemistry

Instruments: I04-Macromolecular Crystallography

Other Facilities: ESRF

crystals-09-00230 (1).pdf